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Updated: Feb 5, 2026
Two-vessel Occlusion Mouse Model of Cerebral Ischemia-reperfusion
Published on: March 1, 2019
miR-29b affects neurocyte apoptosis by targeting MCL-1 during cerebral ischemia/reperfusion injury
Zhi Huang1,2, Lu Lu3, Tianpeng Jiang2
1Department of Intervention, The Affiliated Baiyun Hospital of Guizhou Medical University, Guiyang, Guizhou 550002, P.R. China.
Abstract:
The present study aimed to determine whether an miRNA (miR)-29b inhibitor protected against cerebral ischemia/reperfusion (I/R) injury in vitro and to investigate the underlying mechanisms. As a model for induced cerebral IR injury, N2a cells were exposed to an oxygen-glucose deprivation/reoxygenation (OGD/R) environment. Using this model, it was demonstrated that miR-29b was significantly upregulated compared with cells in a normal environment. The interactions between miR-29b and myeloid cell leukemia sequence (MCL)-1 were then investigated using dual-luciferase assays, revealing a strong regulation of MCL-1 through the 3'untranslated region. Using the OGD/R model, the present study additionally examined the effects of miR-29b and miR-29b inhibitor on cell viability and apoptosis using Cell Counting kit 8 and flow cytometry assays, respectively. miR-29b transfection led to increased N2a cell apoptosis and reduced cell viability under an OGD/R environment. However, this effect was reversed by the miR-29b inhibitor. Finally, the effects of miR-29b on the expression of several Wnt-associating proteins were examined. It was observed that B cell lymphoma-2 was inhibited by miR-29b, as was MCL-1, whereas caspase-3 expression was promoted. The miR-29b inhibitor demonstrated the opposite effect. Overall, miR-29b promoted neurocyte apoptosis by targeting MCL-1 during cerebral I/R injury. The results of the present study suggest a potential novel therapeutic target for the treatment of ischemic stroke.
Insights
MicroRNA 29b (miR-29b) promotes neurocyte apoptosis during cerebral ischemia/reperfusion injury by targeting MCL-1. An miR-29b inhibitor protected against this injury, suggesting a potential therapeutic target for ischemic stroke.
Area of Science:
- Neuroscience
- Molecular Biology
- Biochemistry
Background:
- Cerebral ischemia/reperfusion (I/R) injury is a significant cause of neurological damage.
- MicroRNAs (miRNAs) play crucial roles in regulating cellular processes, including apoptosis and response to injury.
- The specific role of miR-29b in cerebral I/R injury requires further elucidation.
Purpose of the Study:
- To determine if a miR-29b inhibitor protects against cerebral I/R injury *in vitro*.
- To investigate the underlying molecular mechanisms of miR-29b's action in cerebral I/R injury.
- To explore miR-29b's potential as a therapeutic target for ischemic stroke.
Main Methods:
- Establishment of an oxygen-glucose deprivation/reoxygenation (OGD/R) model using N2a cells to mimic cerebral I/R injury.
- Dual-luciferase assays to investigate the interaction between miR-29b and myeloid cell leukemia sequence (MCL)-1.
- Cell Counting kit-8 and flow cytometry assays to assess cell viability and apoptosis.
- Western blot analysis to examine the expression of apoptosis-related proteins.
Main Results:
- miR-29b was significantly upregulated in N2a cells under OGD/R conditions.
- miR-29b directly targets and regulates MCL-1 expression.
- miR-29b transfection exacerbated N2a cell apoptosis and reduced viability under OGD/R.
- Treatment with a miR-29b inhibitor reversed these detrimental effects, improving cell viability and reducing apoptosis.
- miR-29b inhibited BCL-2 expression, promoted caspase-3 activation, and targeted MCL-1, while the inhibitor showed opposite effects.
Conclusions:
- miR-29b promotes neurocyte apoptosis and exacerbates cerebral I/R injury by targeting MCL-1.
- Inhibition of miR-29b demonstrates a protective effect against cerebral I/R injury *in vitro*.
- miR-29b represents a potential therapeutic target for the treatment of ischemic stroke.
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